Abstract: SA-PO332

Epstein-Barr Virus Induced Epithelial Mesenchymal Transition in HK2 Cells

Session Information

Category: Chronic Kidney Disease (Non-Dialysis)

  • 308 CKD: Mechanisms of Tubulointerstitial Fibrosis

Authors

  • Han, Sang Youb, Inje University, GoYang, Korea (the Republic of)
  • Park, Seok ju, Inje University, GoYang, Korea (the Republic of)
  • Hur, Daeyoung, Inje University College of medicine , Busan, Korea (the Republic of)
Background

Epithelial mesenchymal transition (EMT) represents conversion of epithelial cells into mesenchymal phenotype. It is an important mechanism in tissue fibrosis including kidney. Epstein-Barr virus (EBV), which is well known cause of acute febrile illness and lymphoproliferative diseases, is reported to induce tissue fibrosis such as lung, skin, and liver. However, it is not reported its association with renal fibrosis. We tested whether EBV could induce EMT in renal epithelial cells.

Methods

HK2 cells were incubated with EBV. HK2 cells changed their shape from cobble stone appearance into spindle shape. After confirmation of mesenchymal changes of HK2 cells, we evaluated E-cadherin, ZO-1, and beta-catenin for epithelial marker, N-cadherin, vimentin, TGF-beta for mesenchymal marker, MCP-1, IL8, TNF-alpha, IL18 for inflammatory changes, and Slug, Snail, V-Set And Immunoglobulin Domain Containing 4 (VSIG4) and pSTAT3 for EMT marker. All markers were tested using real-time PCR for mRNA or western blotting for protein expression.

Results

The expressions of E-cadherin, ZO-1, and beta-catenin were decreased and N-cadherin, vimentin, TGF-beta mesenchymal markers were increased after EBV exposure. And expressions of Slug, Snail, and pSTAT3 were also increased. And inflammatory markers such as MCP-1, IL8, TNF-alpha, IL18 were increased.
To confirm whether these changes were due to EBV transfection or secretory proteins from EBV, the HK2 cells were stimulated with latent membrane protein 1 (LMP-1), which is representative EBV-related protein. LMP-1 downregulated the expression of E-cadherin, and upregulated those of vimentin and VSIG4. These findings suggested that EMT was induced by LMP-1 alone.
To test VSIG4 involvement in LMP1-induced EMT, VSIG4 siRNA were treated in LMP1-stimulated HK2. The expression of E-cadherin and vimentin were reversed. The expression of E-cadherin was increased and that of vimentin was decreased.

Conclusion

In conclusion, EMT was induced by EBV-associated protein, LMP1, not by EBV in itself. The LMP1-induced EMT was related with VSIG4 changes. This result provided the possibility of EBV-related EMT in renal fibrosis.